Little information exists on the environmental exposures to low levels of organometallic manganese (a principal combustion product from manganese-containing fuel additives) and public health. This study was undertaken to establish biological and environmental levels of manganese among first-grade schoolchildren in the South African cities of Johannesburg and Cape Town. The study was undertaken subsequent to partial introduction of the manganese-containing fuel additive methylcyclopentadienyl manganese tricarbonyl (MMT) to South African petrol in the Johannesburg region only, about 24 months prior to the commencement of the study, and in anticipation of possible future increases in MMT use in the rest of the country. This study involved the measurement of manganese concentrations in the blood of the total number of 814 of grade one schoolchildren (430 and 384 in Cape Town and Johannesburg, respectively), and in water supplies, soil, and classroom dust at a total number of 21 participating schools. The results indicated higher concentrations of manganese in school soil (P=0.0007) and dust (P=0.0071) samples from Johannesburg relative to Cape Town. Similarly, the mean blood manganese concentration in Johannesburg study subjects (9.80 microg/L, SD 3.59) was significantly higher than that in Cape Town study subjects (6.74 microg/L, SD 3.47), after allowing for the clustering effect within schools and adjusting for the confounding effect of population group (P<0.0001). The blood manganese levels of 4.2% and 12.5% of children in Cape Town and Johannesburg, respectively, equaled or exceeded 14 microg/L, the upper normal reference value specified by the Agency for Toxic Substances Disease Registry (ATSDR, 2000). Importantly, levels of manganese in blood were found to be significantly associated with concentrations of manganese in classroom dust at schools.